Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
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Expression and Characterization of Human Immunodeficiency Virus-1 Oligomerized gp140 Protein in Mammalian Cells

포유동물 세포에서 Human Immunodeficiency Virus-1의 Oligomeric gp140 단백의 발현 및 특성

  • Kim, Eun-Ok (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Eun (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Hyun-Soo (College of Veterinary Medicine, Chungnam National University) ;
  • Shin, Kwang-Soon (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Chul-Joong (College of Veterinary Medicine, Chungnam National University)
  • 김은옥 (충남대학교 수의과대학) ;
  • 김은 (충남대학교 수의과대학) ;
  • 김현수 (충남대학교 수의과대학) ;
  • 신광순 (충남대학교 수의과대학) ;
  • 김철중 (충남대학교 수의과대학)
  • Accepted : 2002.02.01
  • Published : 2002.03.25
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Abstract

The envelope glycoprotein of HIV-1 forms an oligomeric complex resulting in playing a role to induce neutralizing antibody and cell-mediate immune responses. The oligomer exists as a trimer of gp120-gp41 heterodimer which mediates HIV-1 attachment and fusion. We made a cDNA clone of gp140 consisting of gp120 and ectodomain of gp41 from the primary African isolate. To express the oligomeric gp140 in mammalian cells, we adopted the Semliki Forest virus (SFV) based expression system. The oligomeric gp140 in the secretory form was expressed and purified from the cell culture supernatant and characterized. The antibody inducing activity of the purified gp140 was also examined in mice inoculation.

HIV-1의 envelope glycoprotein은 중화항체에 의한 체액성 면역반응의 중요한 target으로 surface glycoprotein인 gp120과 transmembrane glycoprotein인 gp41로 이루어져 있다. gp120과 gp41의 ectodomain으로 이루어진 gp140 유전자를 PCR의 방법으로 증폭하고 Semliki Forest virus(SFV) 유래 expression system을 이용하여 mammalian 세포에서 발현하였다. 발현된 gp140은 natural HIV-1에서와 같이 oligomer를 형성하였다. 발현된 gp140을 정제하여 BALB/c 마우스에 접종하여 항체가 형성되었음을 확인하였다.

Keywords

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